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2 ATP + L-valine + 2 L-Baa = 2 ADP + 2 phosphate + L-Val-(L-Baa)2
3 ATP + 3 L-valine + L-Zaa = 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-amino-acid-L-Zaa
ATP + 2 L-valine + L-Yaa = 2 ADP + 2 phosphate + L-Val-L-Val-L-Yaa
n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n
2 ATP + L-valine + 2 L-Baa = 2 ADP + 2 phosphate + L-Val-(L-Baa)2

Baa: Leu, Ile, Met
2 ATP + L-valine + 2 L-Baa = 2 ADP + 2 phosphate + L-Val-(L-Baa)2
Baa: Leu, Ile, Met
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2 ATP + L-valine + 2 L-Baa = 2 ADP + 2 phosphate + L-Val-(L-Baa)2
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3 ATP + 3 L-valine + L-Zaa = 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-amino-acid-L-Zaa

Zaa: Arg, His, Gln, Asn or Phe
3 ATP + 3 L-valine + L-Zaa = 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-amino-acid-L-Zaa
Zaa: Arg, His, Gln, Asn or Phe
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3 ATP + 3 L-valine + L-Zaa = 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-amino-acid-L-Zaa
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ATP + 2 L-valine + L-Yaa = 2 ADP + 2 phosphate + L-Val-L-Val-L-Yaa

Yaa: Arg, Lys, His, Gln, Asn, Ala, Ser, Thr, Gly, Leu, Ile, Met, Phe or Trp
ATP + 2 L-valine + L-Yaa = 2 ADP + 2 phosphate + L-Val-L-Val-L-Yaa
Yaa: Arg, Lys, His, Gln, Asn, Ala, Ser, Thr, Gly, Leu, Ile, Met, Phe or Trp
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ATP + 2 L-valine + L-Yaa = 2 ADP + 2 phosphate + L-Val-L-Val-L-Yaa
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n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n

Xaa: Val, Leu, or Met. n = 2-5
n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n
the enzyme catalyzes the synthesis of oligopeptides from unprotected L-amino acids in an ATP-dependent manner, showing synthesis of various oligopeptides consisting of two to five amino acids, e.g. synthesis of Val-Val-Val tripeptide by the enzyme RizB. The enzyme shows highest activity with Val, Ile, Leu, and Met as substrates. RizB shows specificity toward N-terminal substrates, but is relaxed in the specificity for the C-terminus
n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n
Xaa: Val, Leu, or Met. n = 2-5
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n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n
the enzyme catalyzes the synthesis of oligopeptides from unprotected L-amino acids in an ATP-dependent manner, showing synthesis of various oligopeptides consisting of two to five amino acids, e.g. synthesis of Val-Val-Val tripeptide by the enzyme RizB. The enzyme shows highest activity with Val, Ile, Leu, and Met as substrates. RizB shows specificity toward N-terminal substrates, but is relaxed in the specificity for the C-terminus
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n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n
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2 ATP + 2 L-Val + Gly
2 ADP + 2 phosphate + L-Val-L-Val-Gly
Substrates: -
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2 ATP + 2 L-Val + L-Ala
2 ADP + 2 phosphate + L-Val-L-Val-L-Ala
Substrates: -
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2 ATP + 2 L-Val + L-Arg
2 ADP + 2 phosphate + L-Val-L-Val-L-Arg
Substrates: -
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2 ATP + 2 L-Val + L-Asn
2 ADP + 2 phosphate + L-Val-L-Val-L-Asn
Substrates: -
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2 ATP + 2 L-Val + L-Gln
2 ADP + 2 phosphate + L-Val-L-Val-L-Gln
Substrates: -
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2 ATP + 2 L-Val + L-His
2 ADP + 2 phosphate + L-Val-L-Val-L-His
Substrates: -
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2 ATP + 2 L-Val + L-Ile
2 ADP + 2 phosphate + L-Val-L-Val-L-Ile
Substrates: -
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2 ATP + 2 L-Val + L-Leu
2 ADP + 2 phosphate + L-Val-L-Val-L-Leu
Substrates: -
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2 ATP + 2 L-Val + L-Lys
2 ADP + 2 phosphate + L-Val-L-Val-L-Lys
Substrates: -
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2 ATP + 2 L-Val + L-Met
2 ADP + 2 phosphate + L-Val-L-Val-L-Met
Substrates: -
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2 ATP + 2 L-Val + L-Phe
2 ADP + 2 phosphate + L-Val-L-Val-L-Phe
Substrates: -
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2 ATP + 2 L-Val + L-Ser
2 ADP + 2 phosphate + L-Val-L-Val-L-Ser
Substrates: -
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2 ATP + 2 L-Val + L-Thr
2 ADP + 2 phosphate + L-Val-L-Val-L-Thr
Substrates: -
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2 ATP + 2 L-Val + L-Trp
2 ADP + 2 phosphate + L-Val-L-Val-L-Trp
Substrates: -
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2 ATP + 3 L-leucine
2 ADP + 2 phosphate + L-Leu-L-Leu-L-Leu
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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2 ATP + 3 L-methionine
2 ADP + 2 phosphate + L-Met-L-Met-L-Met
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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2 ATP + 3 L-valine
2 ADP + 2 phosphate + L-Val-L-Val-L-Val
2 ATP + L-Val + 2 L-Leu
2 ADP + 2 phosphate + L-Val-L-Leu-L-Leu
Substrates: -
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2 ATP + L-Val + 2 L-Met
2 ADP + 2 phosphate + L-Met-L-Met-L-Met
Substrates: -
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2 ATP + L-Val + L-Arg-L-Ser
2 ADP + phosphate + L-Val-L-Arg-L-Ser
Substrates: no homopeptides of Val-Val and Val-Val-Val are detected. This suggests that the enzyme preferentially uses dipeptides as the C-terminal substrate
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2 ATP + L-Val + L-Val + L-Arg-L-Ser
2 ADP + 2 phosphate + L-Val-L-Val-L-Arg-L-Ser
Substrates: no homopeptides of Val-Val and Val-Val-Val are detected. This suggests that the enzyme preferentially uses dipeptides as the C-terminal substrate
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2 ATP + Val + 2 L-Ile
2 ADP + 2 phosphate + L-Val-L-Ile-L-Ile
Substrates: -
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3 ATP + 3 L-Val + L-Arg
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Arg
Substrates: -
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3 ATP + 3 L-Val + L-Gln
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Gln
Substrates: -
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3 ATP + 3 L-Val + L-His
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-His
Substrates: -
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3 ATP + 3 L-Val + L-Phe
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Phe
Substrates: -
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3 ATP + 4 L-leucine
3 ADP + 3 phosphate + L-Leu-L-Leu-L-Leu-L-Leu
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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3 ATP + 4 L-methionine
3 ADP + 3 phosphate + L-Met-L-Met-L-Met-L-Met
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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3 ATP + 4 L-valine
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Val
4 ATP + 5 L-leucine
4 ADP + 4 phosphate + L-Leu-L-Leu-L-Leu-L-Leu-L-Leu
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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4 ATP + 5 L-methionine
4 ADP + 4 phosphate + L-Met-L-Met-L-Met-L-Met-L-Met
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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4 ATP + 5 L-valine
4 ADP + 4 phosphate + L-Val-L-Val-L-Val-L-Val-L-Val
ATP + 2 L-leucine
ADP + phosphate + L-Leu-L-Leu
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + 2 L-methionine
ADP + phosphate + L-Met-L-Met
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + 2 L-valine
ADP + phosphate + L-Val-L-Val
ATP + L-Val + L-Arg
ADP + phosphate + L-Val-L-Arg
Substrates: -
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ATP + L-Val + L-Arg-L-Ser
ADP + phosphate + L-Val-L-Arg-L-Ser + L-Val-L-Val-L-Arg-L-Ser
Substrates: no homopeptides of Val-Val and Val-Val-Val are detected. The enzyme preferentially uses dipeptides as the C-terminal substrates
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ATP + L-Val + L-Arg-NHOH
ADP + phosphate + L-Val-L-Arg-NHOH + L-Val-L-Val-L-Arg-NHOH
Substrates: -
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ATP + L-Val + L-Phe-NHOH
ADP + phosphate + L-Val-L-Phe-NHOH + L-Val-L-Val-L-Phe-NHOH
Substrates: -
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ATP + L-valine + L-Val-L-Val
ADP + phosphate + L-Val-L-Val-L-Val
ATP + L-valine + L-Val-L-Val-L-Val
ADP + phosphate + L-Val-L-Val-L-Val-L-Val
additional information
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2 ATP + 3 L-valine

2 ADP + 2 phosphate + L-Val-L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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2 ATP + 3 L-valine
2 ADP + 2 phosphate + L-Val-L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to four amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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2 ATP + 3 L-valine
2 ADP + 2 phosphate + L-Val-L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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2 ATP + 3 L-valine
2 ADP + 2 phosphate + L-Val-L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to four amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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3 ATP + 4 L-valine

3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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3 ATP + 4 L-valine
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to four amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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3 ATP + 4 L-valine
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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3 ATP + 4 L-valine
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to four amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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4 ATP + 5 L-valine

4 ADP + 4 phosphate + L-Val-L-Val-L-Val-L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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4 ATP + 5 L-valine
4 ADP + 4 phosphate + L-Val-L-Val-L-Val-L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + 2 L-valine

ADP + phosphate + L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + 2 L-valine
ADP + phosphate + L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to four amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + 2 L-valine
ADP + phosphate + L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + 2 L-valine
ADP + phosphate + L-Val-L-Val
Substrates: the enzyme synthesizes homo-oligomers of two to four amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + L-valine + L-Val-L-Val

ADP + phosphate + L-Val-L-Val-L-Val
Substrates: -
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ATP + L-valine + L-Val-L-Val
ADP + phosphate + L-Val-L-Val-L-Val
Substrates: -
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ATP + L-valine + L-Val-L-Val-L-Val

ADP + phosphate + L-Val-L-Val-L-Val-L-Val
Substrates: -
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ATP + L-valine + L-Val-L-Val-L-Val
ADP + phosphate + L-Val-L-Val-L-Val-L-Val
Substrates: -
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additional information

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Substrates: dipeptides, tripeptides, tetrapeptides and pentapeptides are synthesized in the reaction mixtures containing Val, Leu, or Met as substrate. When Ile is used, the longest peptides are tetrapeptides. No peptides are detected when the other 15 amino acids are used as substrates. When Leu, Ile, or Met is used as substrate with Val, heteropeptides containing two residues of Leu, Ile, and Met are detected. No heteropeptides containing Tyr. Pro, Cys, Asp or Glu are detected. When other amino acids that are not usable for homopeptide synthesis are used as substrate, heteropeptides containing only one amino acid residue of these amino acids are detected. Substrate recognition at the N-terminus influences the resulting heteropeptide sequence. The enzyme does not participate in peptide-to-peptide ligation. Very low activity with D-amino acids
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additional information
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Substrates: the enzyme catalyzes the synthesis of oligopeptides from unprotected L-amino acids in an ATP-dependent manner. Synthesis of various oligopeptides consisting of two to five amino acids, e.g. synthesis of Val-Val-Val tripeptide by the enzyme RizB. The enzyme shows highest activity with Val, Ile, Leu, and Met as substrates. In the reaction with Val, tetramers are the longest products synthesized, trimer are formed with Co2+. RizB shows specificity toward N-terminal substrates, but is relaxed in the specificity for the C-terminus, mass spectrometric reaction analysis, overview
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